Microplastics in retail shellfish from a seafood market in eastern Thailand: Occurrence and risks to human food safety.

This study aimed to investigate the presence of microplastics in three economically essential shellfish species: green mussels, cockles and spotted babylon. The average abundance of microplastics ranged from 2.41 to 2.84 particles/g wet weight. The predominant shape was fiber, with colors ranging from black-grey to transparent. The size of the microplastics discovered was <1.0 mm. Polystyrene and polyethylene were the most detected types in mussels and cockles, while linen was the predominant type in spotted babylon. The Thai population's estimated annual intake (EAI) of microplastics through shellfish consumption ranged from 20.23 to 1178.42 particles/person/year. The potential human health risks were evaluated using the polymer hazard index (PHI), which led to risk categories III-IV. These findings, along with others from the literature, indicate that shellfish consumption may pose risks to human health, depending on the species consumed and the origin of the specimens.

Binary blends of normal corn starch and cow cockle starch for the slow-thickening behavior upon pasting.

Corn starch with slow thickening property may facilitate more efficient heat transfer and safety of corn starch-thickened foods. Partial substitution of normal corn starch (NCS) with slow-pasting behavior of cow cockle starch (CCS) was hypothesized to impart binary starch blend with slow-thickening effect during hydrothermal heating. To test hypothesis, a series of starch blend dispersions (with weight ratios of CCS to NCS = 75:25, 50:50, 25:75) were prepared at various starch concentrations (6 %, 8 %, 10 %, and 12 %) and subjected to the Rapid Viscosity Analysis (RVA). RVA viscographs of starch blends were compared with that of NCS, suggesting that nearly all starch blends at various concentrations showed longer time span of pasting and lower pasting rate. Although CCS and NCS blend gels exhibited lower Young's modulus and hardness based on textural profile analysis, the sensory panels revealed that 6 % and 8 % starch blend gels (with weight ratio of CCS to NCS = 25:75) showed the mouthfeel analogous to NCS gel. These findings highlight a viable non-chemical modification strategy that enables binary blends of CCS and NCS as a novel gelling agent with slow-pasting property and may aid in safety and high-quality processing of hydrogel foods.

The spatial distribution and physico-chemical characteristic of microplastics in the sediment and cockle (Anadara granosa) from the coastal waters of East Java, Indonesia, and the health hazards associated with cockle consumption.

This study evaluated microplastic (MP) abundances and physico-chemical characteristics in sediments and Anadara granosa along the East Java coast and their health implications. Fibers (74 %) dominated sediment MPs at south coast, while fragments (49-61 %) dominated north coast. Fiber (43-52 %) is the predominant MP in cockle tissues in all locations. Most MP in sediments (31-47 %) and cockle tissues (41-49 %) is black. The majority of microplastics (100-1500 µm) are found in sediment (73-90 %), and cockles (77-79 %). Very weak correlations found between the amount of MP and the length of the cockle shell. However, Spearman correlation shows that as the amount of MP in sediment increases, so does the amount of MP in cockle tissue. Each year, individuals of varying ages consume an average of 20,800 to 156,000 MP items. Cockles contain plasticizer components and microplastic polymers which are classified from II to V regarding of hazard levels, with V being the most hazardous.

Reproductive consequences of thermal stress-induced bleaching in the giant clam Tridacna crocea.

Reproduction is a fundamental process necessary for maintaining a population. However, reproductive processes are sensitive to thermal stress which can cause bleaching in reef organisms such as corals and giant clams. Here we examined the phototrophic and physiological performances, particularly the reproductive processes, in Tridacna crocea during bleaching and recovery periods. Giant clam individuals were induced to bleach at heated treatment (32-33 °C) for 16 days and allowed to recover at 28-29 °C for 35 days. The control giant clams were kept at 28-29 °C. Heated giant clams showed lower phototrophic performances (Fv/Fm and photosynthesis), but their respiration and survival were similar to control giant clams. The gonadosomatic index (GSI) was lower, and the proportion of regressive eggs (i.e., eggs that are no longer viable) was higher in heated than in control giant clams. However, heated giant clams were able to maintain their egg size. In addition, T. crocea showed recovery of phototrophic potential and color of mantle but not of their reproductive output after a month of recovery. Our results indicate that bleaching reduces the reproductive output in giant clams by disrupting their gametogenesis, such as through egg resorption, but giant clams showed potential reproductive strategy, through maintenance of their egg size, to ensure the quality of their offspring. Furthermore, one month of recovery is not sufficient to restore the normal reproductive processes in T. crocea, which may delay their population recovery after a bleaching disturbance.

Oxidative response to accumulation of trace metals in tissue of two bivalves, the Asian green mussel Perna viridis and the blood cockle Tegillarca granosa, living in Pattani Bay, Thailand.

OBJECTIVE: Using bivalves to indicate aquatic pollutants was favorable for discerning the negative effects of high levels of metal accumulation in tissue. We investigated the correlation between trace metal accumulation and the tissue oxidative response of two bivalves. METHODS: The Asian green mussel Perna viridis and the blood cockle Tegillarca granosa were sampled along with seawater and sediments from three locations around Pattani Bay, Thailand. Accumulation of nine trace metals (cadmium, cobalt, copper, chromium, nickel, manganese, iron, zinc, and lead) in seawater, sediments, and tissue and the oxidative tissue response were evaluated. Metal bioaccumulation factor, biota-sediment accumulation factor, and histopathology were also indicated. RESULT: The present study found that P. viridis and T. granosa were macroconcentrators and bioaccumulative of cadmium, and their tissue accumulation of cadmium was strongly related to lipid peroxidation activation. Perna viridis exhibited a higher oxidative response than T. granosa, as indicated by malondialdehyde, catalase, and reduced glutathione levels. CONCLUSION: The present study indicated that P. viridis and T. granosa were macroconcentrators and bioaccumulative of cadmium, and their tissue accumulation of cadmium was strongly related to lipid peroxidation activation. Research has shown discernible negative effects of a high level of metal accumulation in tissue, and deformed and damaged tissues were present in the gills, digestive glands, intestines, and feet of P. viridis and T. granosa.

A cockle-induced bioturbation model and its impact on sediment erodibility: A meta-analysis.

Modelling the dynamics of an estuary and the evolution of its morphology requires a process-based description not only of the physical processes, but also of the influence of benthic fauna on sediment characteristics at ecosystem scale. A meta-analysis was tested as an approach for modelling the effect of bioturbation exerted by the cockle Cerastoderma edule on sediment erodibility. Six different erosion flume datasets were collected to ensure a broad range of experimental conditions including bed shear stress, population characteristics, and sediment composition. First, a model was built to describe the biogenic fluff layer created by C. edule activity in relation to (i) bioturbation activity using the population metabolic rate [mW·m-2] as a proxy for faunal metabolic energy, and (ii) the silt content [%] of the sediment. Second, different erosion models were compared by testing parameterization steps incorporating both erosion of the fluff layer and/or mass erosion of the sediment bed. Structural differences in the flumes and in the preparation of samples in the six different datasets makes it difficult to propose a single model that satisfactorily simulates all the data and encompasses both types of subsequent erosion, that of the fluff layer and that of the underlying consolidated bed. However, a generic model is proposed for the surficial fluff layer erosion covering a moderate range of bed shear stress (<1 Pa). This study shows that including several datasets covering a wide range of environmental conditions is a key to the robustness of this model, and that new insights can be gained by integrating the complexity of sediment features. We expect that this two-part model can be used in broad contexts in terms of cockle populations, estuarine habitats, and climatic conditions and can combined with various hydro-morpho-sedimentary models that include these biological effects.

Trace elements assessment in Cerastoderma glaucum from port areas in the Tunisian Mediterranean coast: The influence of parasites on bioaccumulation.

In the present study, the seasonal concentration of seven trace elements was investigated in sediment and the cockle Cerastoderma glaucum tissues from two port areas in the North-East and South of Tunisia comparing cockles non-parasitized (NP) and parasitized (P) with digenean parasites. Elements concentration in sediments analyzed in both sites revealed that Zinc (Zn), Chromium (Cr), and Lead (Pb) were the most abundant ones, while Cadmium (Cd) and Mercury (Hg) were less abundant. The bioaccumulation of trace metals and Arsenic (As) in the tissues of cockles seems to be modulated by both the infection state and the parasite species. The relationship between bioaccumulation of metals and As, trematode species and abiotic parameters showed that the availability of certain metals for uptake by P cockles of both sites was influenced by the salinity and temperature of the water. Our results corroborate the possibility of using digenean infecting bivalves in biomonitoring aquatic ecosystems.

Toxicity of two pesticides in binary mixture on survival and enzymatic response of Cerastoderma edule - The warming influence.

Pesticide application increased by about 1 million tonnes in the last 3 decades. Pesticides' overuse, coupled with the need for several pesticides to control different pests in the same crop, and its application many times per year, results in dangerous chemical cocktails that enter in aquatic systems, with impacts to the ecosystems and its communities. Climatic changes are currently another great concern, is predicted by the end of the 21st century, the earth's surface temperature will increase by about 4 °C. Bivalve species are reported as essential to the ecosystems' balance. However, they are also indicated as the organisms that will suffer the most serious effects of the temperature increase. So, this work intends to: a) verify the harm of the sub-lethal concentrations of two worldwide used pesticides, oxyfluorfen and copper (Cu), when combined, to Cerastoderma edule at 15 °C and 20 °C; b) assess the changes in the antioxidant defence system, the activity of the neurological enzyme acetylcholinesterase and the nutritive value of C. edule, after exposure to sub-lethal concentrations of oxyfluorfen and Cu, single and in the mixture, at 15 °C and 20 °C; c) observe the interaction between Cu and oxyfluorfen, considering the different biomarkers. Bivalves were exposed to oxyfluorfen and Cu, single and combined, for 96 h, at 15 °C and 20 °C. Results showed lethal effects to the organisms exposed at 20 °C when exposed to the highest binary mixture concentrations. Biochemical effects were observed on the organisms exposed to 15 °C, despite not observing any lethal effects. Briefly, there was a reported increase in oxidative stress and a decrease in protein content, regardless of the increase in the activity of antioxidant enzymes. These results suggest the potentially dangerous effects of the chemicals' mixture combined with the temperature, on this species and its consumers, impacting the trophic chain, and consequently, the community structure and function.

First report of Perkinsus olseni infections in blood cockles Tegillarca granosa on the south coast of Korea.

The protozoan parasite Perkinsus olseni has become a focus of attention since it has been responsible for mass mortalities and economic losses in a wide range of bivalve hosts globally. The P. olseni host range along the south coast of Korea may extend beyond what was previously understood, and blood cockles in the Family Arcidae are also suggested to be potential hosts of P. olseni. In the present study, we applied histology and molecular techniques to identify Perkinsus sp. infections in the blood cockles Tegillarca granosa, which have been commercially exploited on the south coast of Korea for several decades. Histology and molecular techniques, including genus-specific immunofluorescence assay, species-specific fluorescence in situ hybridization, and phylogeny based on the ribosomal DNA internal transcribed spacer region revealed that T. granosa is infected by P. olseni, although the prevalence was low (0.5%). Histology revealed massive hemocyte infiltrations in the mantle, gill, and digestive gland connective tissues, indicating that the infection exerts negative impacts on the host cockles.

Trematode genetic patterns at host individual and population scales provide insights about infection mechanisms.

Multiple parasites can infect a single host, creating a dynamic environment where each parasite must compete over host resources. Such interactions can cause greater harm to the host than single infections and can also have negative consequences for the parasites themselves. In their first intermediate hosts, trematodes multiply asexually and can eventually reach up to 20% of the host's biomass. In most species, it is unclear whether this biomass results from a single infection or co-infection by 2 or more infective stages (miracidia), the latter being more likely a priori in areas where prevalence of infection is high. Using as model system the trematode Bucephalus minimus and its first intermediate host cockles, we examined the genetic diversity of the cytochrome c oxidase subunit I region in B. minimus from 3 distinct geographical areas and performed a phylogeographic study of B. minimus populations along the Northeast Atlantic coast. Within localities, the high genetic variability found across trematodes infecting different individual cockles, compared to the absence of variability within the same host, suggests that infections could be generally originating from a single miracidium. On a large spatial scale, we uncovered significant population structure of B. minimus, specifically between the north and south of Bay of Biscay. Although other explanations are possible, we suggest this pattern may be driven by the population structure of the final host.

Long-term survey discloses a shift in the dynamics pattern of an emerging disease of cockles Cerastoderma edule, marteiliosis, and raises hypotheses to explain it.

Drivers of marine disease outbreaks are poorly understood in spite of their growing impact. We present here results from a unique case study examining how cockles Cerastoderma edule have responded to the introduction of the novel protistan Marteilia cochillia, which led in 2012 to cockle fishery collapse in Galician rias. Based on intensive survey for eight years (2011-2019) of two affected shellfish beds, inner and outer in the Ría de Arousa, involving monthly evaluation of cockle health status and estimation of mortality, detailed information is provided of the declining impact of marteiliosis over a wild cockle population with evidence suggesting its increasing resistance. Disease dynamics involved an annual "breaking wave" of prevalence and subsequent cockle mass mortality, causing the near extinction of every recruited cohort. A shift in this pattern, from a severe epidemic towards an endemic profile, was observed in the inner shellfish bed since the cohort that was recruited in 2016, suggesting the hypothesis of increasing marteiliosis resistance through natural selection. Risk factors that may contribute to trigger marteiliosis outbreaks were analysed. Host age and sex did not influence susceptibility to marteiliosis. No clear relationships between environmental conditions (temperature, salinity and upwelling index) or cockle density and disease dynamics were found. Spatial differences in disease dynamics could be due to differences in the abundance of infective stages hypothetically linked to spatial differences in the population dynamics of a putative planktonic intermediate host. All these findings have potential implications for the management of diseased populations.

Somatic evolution of marine transmissible leukemias in the common cockle, Cerastoderma edule.

Transmissible cancers are malignant cell lineages that spread clonally between individuals. Several such cancers, termed bivalve transmissible neoplasia (BTN), induce leukemia-like disease in marine bivalves. This is the case of BTN lineages affecting the common cockle, Cerastoderma edule, which inhabits the Atlantic coasts of Europe and northwest Africa. To investigate the evolution of cockle BTN, we collected 6,854 cockles, diagnosed 390 BTN tumors, generated a reference genome and assessed genomic variation across 61 tumors. Our analyses confirmed the existence of two BTN lineages with hemocytic origins. Mitochondrial variation revealed mitochondrial capture and host co-infection events. Mutational analyses identified lineage-specific signatures, one of which likely reflects DNA alkylation. Cytogenetic and copy number analyses uncovered pervasive genomic instability, with whole-genome duplication, oncogene amplification and alkylation-repair suppression as likely drivers. Satellite DNA distributions suggested ancient clonal origins. Our study illuminates long-term cancer evolution under the sea and reveals tolerance of extreme instability in neoplastic genomes.

Metallic trace element contamination of the giant clam Tridacna maxima in French Polynesia.

This study is a first assessment of Metallic Trace Elements (MTE) contamination of four lagoons in French Polynesia, chosen because their main anthropic uses are potential sources of MTE (military, agriculture, maritime). We used the giant clam Tridacna maxima as a biointegrator of fifteen MTE to assess the environmental footprint of human activities. The study of organotropism showed that not only the kidney but also the gonad and digestive system (GDS) are accumulator organs of interest for ETM biomonitoring. This result confirms that heterotrophic feeding is the pathway of ETM accumulation. Here, T. maxima appeared to be a super-accumulator of Ni (2700 ppm.dw), but also revealed at Tubuai (As, Fe, Mn) and Arutua (Cd, Zn) the environmental footprint of agricultural and maritime activities. The concentration of Cd recorded in Arutua (5 ppm.dw) both in KID and GDS, also underline the need for better control of effluents resulting from human activities.

Decline of Marteilia cochillia in Ría de Arousa may be due to increased resistance in host Cerastoderma edule.

A huge, unprecedented mortality of cockle Cerastoderma edule caused by the protist Marteilia cochillia, which had never before been detected in Galicia (NW Spain), brought on a cockle fishery collapse in the Ría de Arousa (Galicia) in 2012. Since then, the disease dynamic pattern in the shellfish bed of Lombos do Ulla (at the inner area of that ria) involved an overwhelming annual wave of infections and subsequent cockle mass mortality that caused the near extinction of every cohort recruited to that bed. However, a pattern shift was detected among wild cohorts recruiting since 2016, with progressive declines of marteiliosis prevalence and increments in cockle survival. This suggested 2 non-exclusive hypotheses: increasing marteiliosis resistance through natural selection, and reduced abundance and/or virulence of the parasite. A field experiment was performed to assess these hypotheses by comparing marteiliosis prevalence and severity, as well as mortality, in cockles that naturally recruited to this bed in 2017 and 2018 with those of naïve cockles collected from a marteiliosis-free area and transplanted into Lombos do Ulla in 2017 and 2018. Marteiliosis prevalence and cumulative cockle mortality quickly reached very high values among the transplanted cockles, demonstrating that the parasite remained present and virulent in the area. Conversely, marteiliosis prevalence and cockle mortality were much lower in the cockles that recruited to Lombos do Ulla, suggesting increased resistance that may have been driven by natural selection. The young age at which cockles start reproduction and the very high mortality caused by marteiliosis may have enhanced natural selection.

Exploring the feasibility of cryopreserving larvae of the common cockle (Cerastoderma edule) for hatchery production.

The decline of natural populations of the common cockle (Cerastoderma edule) through the European coast is posing a threat to local small-scale fisheries. These declines are primarily attributed to the prevalence of several pathogens and the disseminated neoplasia in cockle populations. The institution of a biobank of cryopreserved larvae could enhance hatchery production and help the restocking. The present work aimed at the development of a cryopreservation protocol for larvae of the common cockle using the mollusk cryopreservation protocols designed in our laboratory. Toxicity bioassays and short-term cryopreservation experiments were performed for protocol optimization according with cellular tolerance. Once settled, the viability of cryopreserved larvae was studied long term. Toxicity tests evidenced high tolerance of larvae against detrimental effects of Cryoprotecting Agents (CPAs). Cryopreservation of 48 h-old D-larva showed a 100% survival when increasing the equilibrium time from 15 to 60 min and using Propylene-Glycol (PG) + 0.4 M Trehalose (TRE) in Filtered Sea Water (FSW) and 60 min of exposure to CPA solution before slow-cooling. However, when cryopreserving the older larvae, the variation in equilibrium times hardly showed any effect but 10% Ethylene-Glycol (EG) + 0.4 M TRE and 60 min of exposure yielded the best relative survivorship (100%). Cryopreservation caused a significant delay on the growth rate of the latest larval stage. However, cryopreserved larvae survived to day 4-6, while 30 ± 12.17% of control larvae developed into pediveliger stage, of which 50% settled and transformed into juvenile cockles. These results demonstrated the role of the cell-type specificity in cryopreservation and highlight the importance of studying potential long-term effects of this tool to ensure the viability of the protocols.

Heterogeneous microgeographic genetic structure of the common cockle (Cerastoderma edule) in the Northeast Atlantic Ocean: biogeographic barriers and environmental factors.

Knowledge of genetic structure at the finest level is essential for the conservation of genetic resources. Despite no visible barriers limiting gene flow, significant genetic structure has been shown in marine species. The common cockle (Cerastoderma edule) is a bivalve of great commercial and ecological value inhabiting the Northeast Atlantic Ocean. Previous population genomics studies demonstrated significant structure both across the Northeast Atlantic, but also within small geographic areas, highlighting the need to investigate fine-scale structuring. Here, we analysed two geographic areas that could represent opposite models of structure for the species: (1) the SW British Isles region, highly fragmented due to biogeographic barriers, and (2) Galicia (NW Spain), a putative homogeneous region. A total of 9250 SNPs genotyped by 2b-RAD on 599 individuals from 22 natural beds were used for the analysis. The entire SNP dataset mostly confirmed previous observations related to genetic diversity and differentiation; however, neutral and divergent SNP outlier datasets enabled disentangling physical barriers from abiotic environmental factors structuring both regions. While Galicia showed a homogeneous structure, the SW British Isles region was split into four reliable genetic regions related to oceanographic features and abiotic factors, such as sea surface salinity and temperature. The information gathered supports specific management policies of cockle resources in SW British and Galician regions also considering their particular socio-economic characteristics; further, these new data will be added to those recently reported in the Northeast Atlantic to define sustainable management actions across the whole distribution range of the species.

Biochemical responses in mice induced by saxitoxins extracted from the cockles Acanthocardia tuberculatum.

Harmful algae blooms have increased in frequency and geographic range in recent decades, and they produce toxins strains such as saxitoxins (STXs). they block voltage-gated sodium channels and can lead to several poisonings and the death of organisms that pose a significant risk to public and environmental health. The study of STXs toxicity has been carried out but little is known about the response of antioxidant enzymes activities to STXs in mice. The purpose of this study was to evaluate biochemical responses and oxidative stress induced by STXs extracted from Acanthocardia tuberculatum. To this end, daily, mice were treated orally for 7 days with sublethal concentrations (10 mg/100 g mouse). The animal's liver was assessed using biomarkers such as activities of catalase (CAT), thiobarbituric acid reactive substances (TBARS), glyceraldehyde-3-phosphate dehydrogenase (GAPDH) and succinate dehydrogenase (SDH). In the blood, plasmatic markers were analysed as glutamic oxalic transaminase (GOT), glutamic pyruvic transaminase (GPT), creatinine phosphokinase (CPK), lactate dehydrogenase (LDH), urea and creatinine. Globally, test toxicity test showed a significant decrease in the weight at 10 mg /100 g mouse, and the results showed an increase of GPT, GOT, CPK, LDH, CAT and TBARS activities and the inhibitory effect of GAPDH activities but creatinine, urea and SDH activities showed no significative difference from the control. We concluded that STXs induce oxidative stress breaking in mice the balance of the defence system and causing oxidations reactions. Moreover, STXs affect energy metabolism in mice, however, renal function in mice is not affected by exposure to STXs.

Assessment of toxic metal pollution in Yueqing Bay and the extent of metal-induced oxidative stress in Tegillarca granosa raised in this water.

Yueqing Bay is an important economic shellfish culture zone in Zhejiang Province, China. However, increased pollution in the water caused by toxic metals has led to the bioaccumulation of toxic metals in cockles such as Tegillarca granosa, and the consequence of toxic metal-associated toxicity in these animals. This study aimed to assess the concentration of toxic metals in the water and sediment in four different sites (Baisha, Qingjiang, Nanyue, and Wengyang) within Yueqing Bay and to evaluate the extent of metal bioaccumulation in T. granosa raised in the aquaculture farms located within the four sites, as well as the changes in biomarkers in T. granosa in response to the metals. The assessment was carried out at two different times of the year, January and July. The water and sediment samples taken from the aquaculture farms in Baisha (S1), Qingjiang (S2) and Nanyue (S3) were found to have a comprehensive toxic metal pollution index (Pc) <1, indicating that these farms were not polluted. However, the water and sediment samples taken from the aquaculture farm in Wengyang (S4) had a Pc between 1 and 2, indicating mild toxic metal pollution. The edible risk assessments (HQ) of T. granosa in all four farms were <1, and therefore, these cockles could be considered safe for human consumption. The toxic metal enrichment in T. granosa exhibited a strong correlation with the toxic metal content in the sediment. In all four farms, CAT and SOD activity levels in the visceral mass of T. granosa were higher than those found in the foot, and a significantly higher level of CAT activity was detected in July compared with January. Similarly, MDA and H2O2 contents in the visceral mass were also higher in July than in January. Tegillarca granosa individuals taken from S4 and S3 farms exhibited significantly higher levels of metallothionein (MT) mRNA and MDA compared with individuals from S1 and S2 farms. Furthermore, the levels of MDA and MT mRNA showed significant positive correlations with Cd, Cr, Hg, and Cu. Elevation of lipid peroxidation in these cockles coincided with increasing levels of endogenous antioxidants. The visceral mass of T. granosa and its MDA level could be used as a tissue indicator and a biochemical marker, respectively, for detecting toxic metal pollution. MT mRNA might also be used as a molecular marker of toxic metal pollution. The integrated biomarker response version 2 (IBRv2) values of the four aquaculture farms in Yueqing Bay showed the order S4 > S3 > S2 > S1, indicating that S4 had the most serious metal-induced stress. Furthermore, the IBRv2 values correlated with the Nemerow composite index (Pc) for all the cockles examined. Thus, as far as the contamination of aquaculture farms in Yueqing Bay by toxic metals is concerned, the aquaculture farm in Wengyang (S4) was mildly contaminated by toxic metals. However, the contamination was relatively low, presenting a low risk for the local population of T. granosa.

Evaluation of the chemical, physical, and biological properties of a newly developed bioceramic cement derived from cockle shells: an in vitro study.

BACKGROUND: Tricalcium silicate is the main component of commercial bioceramic cements that are widely used in endodontic treatment. Calcium carbonate, which is manufactured from limestone, is one of the substrates of tricalcium silicate. To avoid the environmental impact of mining, calcium carbonate can be obtained from biological sources, such as shelled mollusks, one of which is cockle shell. The aim of this study was to evaluate and compare the chemical, physical, and biological properties of a newly developed bioceramic cement derived from cockle shell (BioCement) with those of a commercial tricalcium silicate cement (Biodentine). METHODS: BioCement was prepared from cockle shells and rice husk ash and its chemical composition was determined by X-ray diffraction and X-ray fluorescence spectroscopy. The physical properties were evaluated following the International Organization for Standardization (ISO) 9917-1;2007 and 6876;2012. The pH was tested after 3 h to 8 weeks. The biological properties were assessed using extraction medium from BioCement and Biodentine on human dental pulp cells (hDPCs) in vitro. The 2,3-bis(2-methoxy-4-nitro-5-sulfophenyl)-5[(phenylamino)carbonyl]-2 H-tetrazolium hydroxide assay was used to evaluate cell cytotoxicity following ISO 10993-5;2009. Cell migration was examined using a wound healing assay. Alizarin red staining was performed to detect osteogenic differentiation. The data were tested for a normal distribution. Once confirmed, the physical properties and pH data were analyzed using the independent t-test, and the biological property data were analyzed using one way ANOVA and Tukey's multiple comparisons test at a 5% significance level. RESULTS: The main components of BioCement and Biodentine were calcium and silicon. BioCement's and Biodentine's setting time and compressive strength were not different. The radiopacity of BioCement and Biodentine was 5.00 and 3.92 mmAl, respectively (p < 0.05). BioCement's solubility was significantly higher than Biodentine. Both materials exhibited alkalinity (pH ranged from 9 to 12) and demonstrated > 90% cell viability with cell proliferation. The highest mineralization was found in the BioCement group at 7 days (p < 0.05). CONCLUSIONS: BioCement exhibited acceptable chemical and physical properties and was biocompatible to human dental pulp cells. BioCement promotes pulp cell migration and osteogenic differentiation.

One-year variation in quantity and properties of microplastics in mussels (Mytilus galloprovincialis) and cockles (Cerastoderma edule) from Aveiro lagoon.

As filter feeders, marine bivalves inhabiting estuarine and coastal areas are directly exposed to microplastics (MPs) in water. To assess whether MPs number, and their shape, size, colour, and polymer type present in mussels (Mytilus galloprovincialis) and cockles (Cerastoderma edule) varied over one year, bivalves were collected over the year of 2019 in the lower part of the coastal Aveiro lagoon, Portugal. After extraction from the bivalve's whole-body soft tissues, a subset of the visually inspected particles was randomly separated for identification using the Fourier-transform mid-infrared (FT-MIR) spectroscopy. A fraction of the inspected particles, 26-32% of particles >100 µm, and 59-100% of smaller ones were confirmed as MPs. Concentrations varied within the intervals of 0.77-4.3 items g-1 in mussels and 0.83-5.1 items g-1 in cockles, with the lowest values observed in January. In winter, the accumulation of large-sized fibers was composed of a mixture of plastic types, which contrasted against the most abundant MPs in summer consisting mainly of polyethylene of diverse size classes and shapes. Temperature decrease registered in winter might have triggered a lower filtration rate, resulting in lower MPs concentrations in the whole-soft body tissues of organisms. Different properties of MPs found in bivalves between January-February and August-September appear to reflect changes in the characteristics of MPs available in the Aveiro lagoon.